1.1 Field of the Invention
The present invention is generally directed to the field of carbohydrate analysis, and more particularly to the analysis of carbohydrates derivatized with a visible chromophoric dye by high resolution polyacrylamide gel electrophoresis. The invention also includes methods and kits for analyzing carbohydrates.
1.2 Description of Related Art
Carbohydrates have many important biological functions. Carbohydrates play important roles in development, immunology and cancer metastasis (Shur, 1994; Wright and Morrison, 1997; Kawaguchi, 1996). Glycoproteins are involved in cell stability and adhesion, antibody recognition, and microorganism binding, and also serve as cell-surface markers (Wyss and Wagner, 1996; Hounsell et al., 1996; Parham, 1996).
Thus, there is substantial interest in carbohydrate analysis. However, a technique for carbohydrate analysis that is both inexpensive and easily performed is currently unavailable. High-sensitivity carbohydrate analysis has been performed with gas chromatography (Karlsson et al., 1995; Karlsson et al., 1994; Hansson and Karisson, 1993), liquid chromatography (Starr et al., 1996; Hu, 1995; Weitzhandler et al., 1993), mass spectrometry (Reinhold et al., 1996; Kovacik et al., 1996; Hellerqvist and Sweetman, 1990), nuclear magnetic resonance spectroscopy (Sugahara et al., 1996; Larnkjaer et al., 1995; Pozsgay and Coxon, 1994; Haynes et al., 1992) and capillary electrophoresis (El Rassi and Mechref, 1996; Oefner and Chiesa, 1994; Liu et al., 1991). However, the above methods of carbohydrate analysis require sophisticated instrumentation and highly trained personnel.
A method for carbohydrate analysis that utilizes polyacrylamide gel electrophoresis to separate fluorescently-labeled carbohydrates has been developed (Jackson, 1996; Jackson, 1994a; Jackson, 1994b) and optimized (Klockow et al., 1996; Masada et al., 1996). However, fluorometers for detection and quantification of the modified carbohydrates are not widely available. Thus, a technique for carbohydrate analysis that is both inexpensive and easily performed is currently unavailable.